Derivatives of lincomycin and its analogs and process

ABSTRACT

ALKYL 7-DEOXY-7(S)-OR-A-THIOLINCOSAMINIDES USEFUL AS INTERMEDIATES FOR PREPARING ANTIBACTERIALLY ACTIVE 7-DEOXY7(S)-OR-LINCOMYCINS ARE PREPARED BY OPENING THE AZIRIDINE RING OF ALKYL N-ACYL-6,7-ARIZIDINO-6-DEAMINO-7-DEOXYA-THIOLINCOSAMINIDES BY HYDROLYSIS OR ALCOHOLYSIS.

United States Patent DERIVATIVES 0F L'INCOMYCIN AND ITS ANALOGS ANDPROCESS Brian Bannister, Kalamazoo, Mich., assignor to The UpjohnCompany, Kalamazoo, Mich. N0 Drawing. Filed Apr. 6, 1970, Ser. No.26,119 Int. Cl. C07c 47/18 US. Cl. 260-210 R 28 Claims ABSTRACT OF THEDISCLOSURE Alkyl 7-deoxy-7(S)-OR-a-thiolincosaminides useful asintermediates for preparing antibacterially active 7-deoxy-7(S)-OR-lincomycins are prepared by opening the aziridine ring of alkylN-acyl-6,7-aziridino-6-deamino-7-deoXya-thiolincosaminides by hydrolysisor alcoholysis.

BRIEF DESCRIPTION OF THE INVENTION The invention relates to alkyl7-d6OXy-7(S)OR-ot-thl0- lincosaminides of Formula I and acylates thereofand to a process for making the same HO T la 2i S-Alkyl Alkyl is alkylof not more than 4 carbon atoms, for example, methyl, ethyl, propyl,isopropyl, butyl, sec.-butyl, isobutyl, and tert.-butyl and R ishydrogen or a hydrocarbon radical, advantageously containing not morethan 12 carbon atoms, and hydrocarbon radicals substituted by one ormore hydroxy, alkoxy, or halogen groups, ad- 'vantageously containingnot more than 12 carbon atoms. Suitable hydrocarbon and hydroxy, alkoxy,and halo-substituted hydrocarbon radicals include alkyl, aralkyl,cycloalkyl, aryl, and hydroxy, alkoxy, and halo-substituted alkyl,aralkyl, cycloalkyl, and aryl. Suitable such radicals include methyl,ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl,undecyl, and duodecyl, and the isomeric forms thereof; hydroxyethyl,glyceryl, alkoxyethyl where the alkoxy group contains advantageously notmore than carbon atoms; 2-chloroethyl, 2-bromoethyl, 2- iodoethyl,benzyl, o-, m-, and p-hydroxybenzyl, o-, m-, and p-halobenzyl where halois chlorine, bromine, iodine; 0-, m-, and p-alkoxybenzyl and o-, m-, andp-alkoxyphenethyl where alkoxy is as given above; phenethyl, o-, m-, andp-hydroxyphenethyl, o-, m-, and p-halophenethyl where halo is chlorine,bromine, or iodine; cyclohexyl, cyclopentyl, 3-cyclopentylpropyl,phenyl, 0-, m-, and phydroxyphenyl, o-, m-, and p-halophenyl where halois chlorine, bromine, or iodine, o-, m-, and p-alkoxyphenyl where alkoxyis as given above, 04- and p-naphthyl, aand fl-naphthylmethyl, aandfl-naphthylethyl.

The compounds of Formula I can be obtained in several different ways.All of the processes involve the opening of the aziridine ring of amethyl or ethyl 6,7-aziridino-6- deamino-7-deoxy-a-thiolincosaminide ofthe formula H II 3,702,322 Patented Nov. 7, 1972 AeNH- A01 K 0 2 OAGi I--Alkyl in which Ac is a carboxacyl and Ac, is hydrogen. The carboxacylgroup is then removed by hydrazinolysis in a manner already well knownin the art (See US. 3,179,- 565 to yield alkyl7(S)-O-a1kyl-7-deoXy-a-thiolincosaminide of Formula I.

The compounds of the invention (Formula I) are useful for the samepurposes as methyl a-thiolincosaminide (methyl6-arnino-6,8-dideoxy-l-thio D erythro a D- galacto-octopyranoside,oc-MTL) as disclosed in US. Pat. 3,380,992 and moreover can be acylatedwith trans-1- methyl-4-propyl-L-2-pyrrolidine carboxylic acid and likeacids to form 7-deoxy-7(S)-OR-derivatives of lincomycin and analogsthereof which are useful as antibacterial agents.

In accordance with a preferred embodiment of the invention the startingcompound of Formula II is first allowed to react with acetic anhydrideor like carboxacyl anhydride in methanol to give alkylN-acyl-7-deoxv-7(S methoxy-a-thiolincosaminide (methyl7-O-methyl-6-acylamino-6,8-dideoxy-l-thio-L-threo a D galactooctopyranoside). The reaction proceeds smoothly at room temperaturethough higher temperatures say up to about 30 C. can be used. Any lowertemperature which does not slow the reaction down too much can be used.

The N-acyl group can then be removed by hydrazinolysis in a manneralready well known in the art.

It is sometimes advantageous, however, to fully acetylate the product ofthe methanolysis, that is the alkyl N- acetyl-7-deoxy-7(S)-methoxy ccthiolincosaminide, in order to provide a product which is more easilypurified. When the methanolysis product, of Formula III where Ac ishydrogen, is acylated with acetic anhydride, or like carboxacylanhydride, in pyridine in a manner already well known in the art, theproduct under these conditions has been shown to be a mixture containingmainly the product of Formula III where both Ac and Ac, are acyl groupsand a minor amount of a triacylate in which the 4-Ac is hydrogen.

The reaction mixture can be purified by simple crystallization or simplechromatography and then converted to the desired product (Formula I) byhydrazinolysis or it can be resolved into its components by separationtechniques already well known in the art such as liquid-liquidcountercurrent extraction, for example Craig countercurrent distributionor partition chromatography.

By substitution of the methanol by other hydroxy compounds of theformula ROH wherein R is as given above the corresponding 7-ORderivatives are obtained.

By substituting the methanol by water there are obtained compounds ofFormula HI wherein Ac, and R are hydrogen which on hydrazinolysis yieldalkyl 7-epi-athiolincosaminides of Formula I where R is hydrogen whichon acylation with the appropriate L-2-pyrrolidinecarboxylic acid yield7-epilincomycin and analogs thereof as disclosed in US. Pat. 3,380,992.

III

When the starting compound is N-acylated, it is not necessary, if timepermits, to use acid catalyzed hydrolysis or acid catalyzed alcoholysis.A mild acid condition greatly accelerates the solvolysis, though. Thusit is of advantage to effect the solvolysis under mildly acidicconditions such as are obtained with acetic acid or like carboxylicacids.

In accordance with another preferred embodiment of the invention, theaziridine starting compound of Formula II is acylated with aceticanhydride or like carboxacyl anhydrides in an alkyl carbinol, forexample, ethanol, propanol, and butanol. The reaction takes placereadily at room temperature. Higher or lower temperatures can be usedbut temperatures high enough to cause peracylation should be avoided.Generally speaking temperatures above 30 C. should be avoided. Theresult is acylation at the nitrogen only to give a product of thefollowing formula wherein Ac is hydrogen. On subjecting the resultingcompound to hydrolysis or alcoholysis in the presence of acetic acid, orlike carboxylic acid, with gentle heating, for example, at the refluxtemperature, there is obtained a compound of Formula III wherein Ac isacyl, Ac, is hydrogen, and R is hydrogen or the radical of the alcoholused in the alcoholysis. The resulting product can be processed in waysalready described to give the desired compound (Formula I).

When the alcohol in the latter embodiment is ethanol-- i.e., whereethanol is the alkyl carbinol-both acylation and alcoholysis resultconcomitantly so that half of the product is N-acyl-aziridine of FormulaIV where Ac is hydrogen and the other half is the desired ring-openedproduct of Formula III where Ac, is hydrogen. With other alkyl carbinolslittle or no ring opening results.

In accordance with another preferred embodiment of the invention thestarting aziridine compound of Formula II is peracylated using aceticanhydride or like carboxacyl anhydride in pyridine or other acid bindingagent, to give a compound of Formula IV in which both Ac and Ac arecarboxacyl groups. The resulting product is then sub jected toalcoholysis in acetic acid with heating in a manner already described togive compound of Formula III in which Ac and Ac, are both carboxacyl.This process has the advantage that the resulting product is easilypurified by such techniques as crystallization and absorption orpartition chromatography and is readily converted to the desiredcompound of Formula I by hydrazinolysis in a manner already described.

The starting aziridino compounds of Formula II can be obtained bydehydrohalogenation of methyl 7(8)- chloro-7-deoxy-a-thiolincosaminide(Belgian Pat. 705,427, Apr. 22, 1968; U.S. application 692,727, filedDec. 22, 1967). The dehydrohalogenation is efiected with anhydroussodium carbonate by heating at reflux in dimethylformamide (Belgian Pat.732,352, October 1969; U.S. application 725,531, filed Apr. 30, 1968).

By acylating the compounds of the invention (Formula I) with anL-2-pyrrolidinecarboxylic acid, compounds of Formula III in which Ac isthe acyl of the L-2-pyrrolidinecarboxylic acid and Ac is hydrogen areobtained. When alkyl and R are methyl and the L-2-pyrrolidinecarboxylicacid is trans-l-methyl-4-propyl-L-2-pyrrolidinecarboxylic acid, thecompound is 7(S)-methoxy-7-deoxylincomycin which has antibacterialactivity in the order of six or seven times greater than theantibacterial activity of the corresponding unalkylated compound, i.e.,

7-epilincomycin. It can be utilized for the same purposes as lincomycinbearing in mind that it is several times more active than lincomycin.

The compounds of the invention (Formula I) as well as the acylatesthereof with an L-2-pyrrolidinecarboxylic acid can exist in either thefree base form or in the form of an acid addition salt. These acidaddition salts can be made by neutralizing the free base with theappropriate acid to below about pH 7.0, and advantageously to about pH 2to pH 6. Suitable acids for this purpose include hydrochloric, sulfuric,phosphoric, thiocyanic, fiuosilicic, hexafluoroarsenic,hexafluorophosphoric, acetic, succinic, citric, lactic, maleic, fumaric,pamoic, cholic, palmitic, mucic, camphoric, glutaric, glycolic,phthalic, tartaric, lauric, stearic, salicylic, 3-phenylsalicylic,S-phenylsalicyclic, S-methylglutaric, orthosulfobenzoic,cyclohexanesulfamic, cyclopentanepropionic, 1,2 cyclohexanedicarboxylic,4 cyclohexenecarboxylic, octadecenylsuccinic, octenylsuccinic,methanesulfonic, benzenesulfonic, helianthic, Reineckes,dimethyldithiocarbamic, hexadecylsulfamic, octadecylsulfamic, sorbic,monochloroacetic, undecylenic, 4' hydroxyazobenzene-4-sulfonic,octeyldecylsulfuric, picric, benzoic, cinnamic, and like acids.

The acid-addition salts can be used for the same purposes as the freebase or they can be employed to upgrade the same. For example, the freebase can be con verted to an insoluble salt, such as the picrate, whichcan be subjected to purification procedures, for example, solventextractions and washings, chromatography, fractional liquid-liquidextractions, and crystallization and then used to regenerate the freebase form by treatment with alkali or to make a different salt bymetathesis. Or the free base can be converted to a water-soluble salt,such as the hydrochloride or sulfate and the aqueous solution of thesalt extracted with various water-irnmiscible solvents beforeregenerating the free base form by treatment of the thus-extracted acidsolution or converted to another salt by metathesis.

The free bases can be used as bufiers or as antacids. They react withisocyanates to form urethanes and can be used to modify polyurethaneresins. The thiocyanic acid addition salt when condensed withformaldehyde forms resinous materials useful as pickling inhibitorsaccording to U.S. Pats. 2,425,320 and 2,606,155. The free bases alsomake good vehicles for toxic acids. For example, the fluosilicic acidaddition salts are useful as mothproofing agents according to U.S. Pats.1,915,334 and 2,075,359 and the hexafluoroarsenic acid andhexafiuorophosphoric acid addition salts are useful as parasiticidesaccording to U.S. Pats. 3,122,536 and 3,122,552.

The invention can be more fully understood by reference to the followingexamples in which the solvent ratios are volume to volume and the partsare by weight unless otherwise specified.

EXAMPLE 1.7-Deoxy-7(S)-methoxylincomycin hydrochloride Part A-l: MethylN-acetyl-7-deoxy-7(S)-methoxy-athiolincosaminide OMe AcNH- 4.8 x 94 cm.column of silica gel using 1 MeOHzlO CHC1 as the solvent system. Theweight of the silica was 750 gms. After a forerun of 1000 ml., 50 ml.fractions were collected. Fractions 31-85 were combined, and evaporatedto dryness yielding 3.2 gms. of methyl N-acetyl-7(S)methoxy-7-deoxy-a-thiolincosaminide (VI) as a colorlessamorphous solid, having the molecular weight by mass spectrometry of309, compared with the calculated molecular weight of 309.38.

Part B-l: Methyl 7-deoxy-7 (S)methoxy-a-thiolinosaminide (VII) (Methyl6,8 dideoxy-7-O-methyl-6-amino-lthio-L-threo-u-D-galacto-octopyranoside)OMe NHz-

\l Me VII A solution of 3.2 gms. of methyl7-deoxy-7(S)-methoxy-a-thiolincosaminide (VI) in 25 gms. of hydrazinehydrate was heated under gentle reflux with stirring in an oil-bath at145 C. overnight. The solvent was removed from the colorless solution ascompletely as possible by distillation from an oil-bath at 100 C./15 mm.and finally at high vacuum to give methyl7-deoxy-7(S)-methoxyu-thiolincosaminide as a colorless syrup. The syrupwas chromatographed on 750 gms. of silica gel in a 4.8 x 97 cm. columnusing 1 MeOHzlO CHCl as the solvent system. After 1.4 liter forerun, 50ml. fractions were collected. Fractions 281-600 were pooled andevaporated to dryness yielding 2.06 gms. methyl 7-deoxy-7(S)-methoxya-thiolinocosaminide (VI) which on crystallization fromacetonitrile yielded colorless needles having the followingcharacteristics: M.P. 154155 C. [a] +260 (c., 0.5634, H O).

Analysis.-Calcd. for C H O NS (percent): C, 44.92; H, 7.92; N, 5.24; S,12.00; OMe, 11.61. Found (percent): C, 45.20; H, 7.96; N, 5.08; S,12.19; OMe, 11.86. Mol. wt. calcd.: 267.35. Found (mass spec.): 2.67.

VIII

To a suspension of 2.7 gms. of trans-l-methyl-4-propyl-L-2-pyrrolidinecarboxylic acid hydrochloride in 90 ccs. acetonitrile wasadded with stirring 2.89 gms. of triethylamine. The stirring wascontinued until all of the solid had dissolved; the reaction mixture wasthen cooled in an ice/methanol bath to 5 C., when a precipitate oftriethylamine hydrochloride appeared. There was then added 1.78 gms. ofisobutyl chloroformate dropwise keeping the temperature of the reactionat -5 to 3 C. Additional triethylamine hydrochloride precipitated andstirring was continued at 5 C. for 20 minutes. To the resulting reactionmixture was added 1.74 gms. of methyl 7-deoxy-7(S)-methoxy orthiolincosaminide (VII), dissolved in 10 ccs. of water. As the solidsdissolved, the temperature rose to about 0 C. and stirring was continuedfor 2 hours, without further icing the cooling bath. The solvent wasthen removed on a rotary evaporator at 40 C./l5 mm. to a brown viscousresidue. This was dissolved in dilute hydrochloric acid and the solution(pH 2) extracted twice with chloroform and the combined extracts washedonce with lwater. The aqueous phase containing the wash water wasadjusted to pH 11 with sodium hydroxide (50% aqueous solution),saturated with sodium chloride and extracted 3 times with chloroform.The combined chloroform extracts were dried over anhydrous sodiumsulfate and taken to dryness yielding 1.76 gms. of a tan amorphoussolid. The tan amorphous solid was chromatographed on 750 gms. of silicagel in a 4.8 x 94 cm. column using 1 MeOHzlS CHCl as the solvent system.After 1.3 liters of forerun, 50 ml. fractions were collected. Fractions60 through were pooled and taken to dryness yielding7-deoxy-7(S)-methoxylincomycin free base as an almost colorless syrup.This free base was taken up in dilute aqueous HCl and the resultingsolution filtered and freeze-dried yielding 801.4 mg. of7-deoxy-7(S)methoxylincomycin hydrochloride as a colorless amorphoussolid having the following characteristics: [111 +117 (c., 0.9626, H O).

Analysis.--Calcd. for C H O N S-HCl (percent): C, 49.93; H, 8.16; N,6.13; S, 7.02. Found (corrected for 5.14% H O) (percent): C, 49.44; H,7.99; N, 6.20; S, 6.48.

M01. wt. Calcd. for anhydrous free base: 420.57

Found (Mass spec.): 420

Antibacterial activity: 2 to 4 times lincomycin hydrochlorideAntibacterial spectrum: Same as lincomycin Antibacterial activityagainst mice infected with Staphylo- COCCMS aureus:

CD =7.0 (4.4-11.1) mg./kg.= /z to 1 times the activity of7(S)-chloro-7-deoxylincomycin hydrochloride.

EXAMPLE 2.-ALTERNATIVE METHOD FOR PRO- DUCING METHYL 7-DEOXY-7 (S)METHOXY-a- THIOLINCOSAMINIDE (VII) Part A-2: MethylN-acetyl-6,7aziridino-6-deamino-7- deoxy2,3,4-tri-O-acetyl-a-thiolincosaminide (IX) 0 Ac IX To a solution of 2.0gms. of methyl 6,7-aziridino-6- deamino-7deoxy-a-thiolincosaminide (V)in 20 ccs. of pyridine was added with stirring 10 ccs. of aceticanhydride and the reaction mixture left overnight at room temperature.The volatile material was removed as completely as possible from thereaction mixture on a rotary evaporator at 40 C./7 mm., finally at highvacuum, to a colorless solid. The resulting solid was dissolved inchloroform, stirred with aqueous cadmium chloride to remove thepyridine, filtered and the chloroform layer washed twice with water, anddried over anhydrous sodium sulfate. On removal of the solvent on therotary evaporator at 40 C./7 mm. methylN-acetyl-6,7-aziridino-6-deamino-7-deoxy-2,3,4-tri-O-acetyl athiolincosa'minide (IX) was obtained as a colorless crystalline solid,weight 3.1 gms. Recrystallization from ethyl acetate-Skellysolve B(technical hexane) gave colorless prismatic needles having the followingcharacteristics: M.P. 173.5- C. [01], +222 (c., 0.912, CHClAnalysis.Calcd. for C H O NS (percent): C, 50.61; H, 6 .25; N, 3.47; S,7.95. Found (percent): C, 50.43; H, 6.33; N, 3.41; S, 8.31. M01. wt.calcd.: 403.45. Found (Mass spec.): 403.

7 'Part B-2: Methyl N-acetyl-2,3,4-tri-O=acetyl-7-deoxy- 7 (S'HlfithOXY-u-thlOlllICOSflIIllIlldC (X) A mixture of 5 gms. of methylN-acetyl-2,3,4-O-tri acetyl-6,7-aziridino-6-deamino 7 deoxy cthiolincosaminide (IX) 50 ccs. methanol, and ccs. glacial acetic acidwas heated under gentle reflux in an oil bath at 130 C. for six hours.The solvent was removed from the colorless solution at 40 C./7 mm. on arotary evaporator yielding a pale yellow syrup which crystallized. Thecrystals were taken up in methylene chloride solution, washed withsaturated aqueous sodium bicarbonate, then with water, and then driedover anhydrous sodium sulfate. Removal of the solvent as above gavemethyl N-acetyl-2,3,4- tri-O-acetyl-7(S)-methoxy-7-deoxy-athiolincosaminide (X) as colorless crystals (53.1 gms.). Crystallizationfrom ethyl acetate-Skellysolve B gave fine colorless needles having thefollowing properties: M.P. 235236 C. [a] +205 (c., 0.9952, CHClAnalysis.Calcd. for C H O NS (percent): C, 49.64; H, 6.71; N, 3.22; S,7.36; OMe, 7.13. Found (percent): C, 49.77; H, 6.92; N, 3.65; S, 7.90;OMe, 7.38. M01. wt. calcd.: 435.49. Found (Mass spec.): 435.v

On hydrazinoylsis by the procedure of Part B-l there is obtained methyl7-deoxy-7(S)-methoxy-a-thiolincosaminide (VII).

EXAMPLE 3Modification of Example 1 Part A-S: Methyl Nacetyl-2,3,4tri-O-acetyl-7-deoxy- 7(S) methoxy cc thiolincosaminide (X)and methyl N acetyl 2,3 di O acetyl 7 deoxy-7(S)- methoxy athiolincosaminide (XI) CH; CH;

-0Me OMe .AcNH- AcNH- AcO no I 0Ac y K0110 l l/ SMe \I SMe 0A 0A0 X XITo 26.61 gms. of methyl N-acetyl 7 deoxy-7(S)-methoxy-a-thiolincosaminide (VI) in 100 ccs. of pyridine there was added50 ccs. of acetic anhydride with stirring and the reaction mixtureallowed to stand overnight at room temperature. The volatile materialswere then removed by distillation on a rotary evaporator at 40 C./ 7 mm.and finally under high vacuum. The residue was dissolved in chloroformand washed with saturated aqueous sodium bicarbonate. The aqueous layerwas washed with chloroform and the combined chloroform extracts stirredwith aqueous cadmium chloride to remove the pyridine.

The precipitate was filtered off and washed well with chloroform and thechloroform layer separated, washed twice with water and dried overanhydrous sodium sulfate. On removal of the solvent on a rotaryevaporator at 40 C./ 7 mm. a pale yellow syrup which crystallized onstanding was obtained. On recrystallization from ethyl acetate-Skellysolve B, the product was obtained as small colorless, flattenedneedles, and had the following characteristics: M.P. 245-247 C. [M +202(c., 0.7142, CHCl Analysis.-Calcd. for C H O NS (percent): C, 49.64; H,6.71; N, 3.22; S, 7.36; OMe, 7.13. Found (percent): C, 49.24; H, 6.75;N, 3.34; S, 7.52; OMe, 7.17. M01. wt. calcd.: 435.49. Found (Massspec.): 435.

The above material by Craig countercurrent distribu tion using as asolvent system 1 EtOI-I:l H 011 EtOAczl cyclohexane was shown to contain70% of methyl N- acetyl 2,3,4 tri O acetyl 7 deoXy-7 (S)-methoxya.thiolincosaminide (X) and 30% of methyl N-acetyl- 2,3 di O acetyl 7deoxy-7(S)-methoxy-a-thio lincosaminide (XI). After 500 transfers,fractions from tubes 225-310 were pooled (K value 1.14) and evaporatedto dryness and on recrystallization from ethyl acetate- Skellysolve Bgave methyl N-acetyl-2,3,4tri-O-acetyl-7- deoxy-7(S)-methoxy athiolincosaminide (X) as fine colorless needles, identical with theproduct of Part B-2.

Fractions from tubes -220 (K value 0.59) were pooled and evaporated todryness and on recrystallization from ethyl acetate-Skellysolve B gavemethyl N-acetyl- 2,3 di O acetyl 7deoxy-7(S)-methoxy-a-thiolincosaminide (XI) as colorless chunky needleshaving the following characteristics: M.P. 189-190 C. [a] +275 (c.,1.0188, CHCl Analysis.Calcd. for C H O NS (percent): C, 48.84; H, 6.92;N, 3.56; S, 8.15; OMe, 7.89. Found (percent): C, 48.71; H, 7.11; N,3.93; S, 7.96; OMe, 7.98. M01. wt. calcd.: 393.46. Found (Mass spec.):393.

Part B-3: Acetylation of methyl N-acetyl 2,3-di-O-acetyl- 7-deoxy-7(S)-methoxy-u-thiolincosaminide (XI) To a solution of 200 mg. of methylN-acety1-2,3-di-O- acetyl 7 deoxy 7(S) methoxy oz thiolincosaminide (X1)in 20 ccs. of pyridine was added 10 ccs. of acetic anhydride withstirring and the reaction mixture left at room temperature overnight.The solvent was removed from the colorless reaction solution on arotating evaporator at 40 C./7 mm. finally at 40 C./high vacuum. Thesyrupy residue was dissolved in chloroform, washed with dilute aqueousHCl /2 normal), twice with water, with saturated sodium bicarbonatesolution and twice with water, and dried over anhydrous sodium sulfate.The solvent was then removed on a rotating evaporator at40 C./7 mm.yielding methyl N-acetyl-Z,3,4-tri-O-acetyl-7-deoxy-7(S)-methoxy-tat-thiolincosaminide (X) as a colorless syrup whichcrystallized on standing.

On hydrozinolysis of the products of Part A-3 and B-3, there is obtainedmethyl 7 deoxy 7(S)-methoxy-uthiolincosaminide (VII).

EXAMPLE 4Alternative for Example 1 Part A-4: Methyl N-acetyl6,7-aziridino-6-deamino-7- deoxy-a-thiolincosaminide (XII) To asuspension of 2.3 gms. of methyl 6,7-aziridino- 6 deamino 7 deoxy ozthiolincosaminide (V) in 25 ccs. isopropyl alcohol, there was added withstirring 2.04 gms. acetic anhydride. Most of the solid appeared to gointo solution to be replaced by new solid. The reaction mixture wasstirred overnight at room temperature, then filtered and the residuewashed with isopropyl alcohol and dried in a vacuum oven at 60 C./l5 mm.There was obtained 2.28 gms. of methyl N-acetyl-6,7-aziridino-6-deamino-7-deoxy-a-thiolincosaminide as colorless platelets having thefollowing properties: M.P. C. [M +253 (c., 0.7916, H O).

Analysis.Calcd. for C H O NS (percent) C, 47.63; H, 6.91; N, 5.05; S,11.56. Found (percent): C, 47.57; H, 6.71; N, 5.23; S, 11.29. Mol. wt.calcd.: 277.34. Found (Mass spec.): 277.

Part B-4: Methyl N-acetyl-7-deoxy-7(S)-methoxy-athiolincosaminide (VI)On treating methyl N-acetyl 6,7-azirindino-6-deamino- 7-deoxy athiolincosaminide (XII) with methanol and acetic acid under reflux,there is obtained methyl N-acetyl- 7 deoxy 7(S) methoxy athiolincosaminide (VI) identical with the product of Part A-l. 4

9 EXAMPLE 5.7-Deoxy-7 (S)-ethoxylincomycin Hydrochloride Part A-5:Methyl N-acetyl-7-deoxy-7(S)-ethoxy-a thiolincosaminide (XIII) AcNH----SMe 0H XIII On treating the methyl N-acetyl-6,7-aziridino-6deamino-7-deoxy-a-thiolincosaminide (XII) with ethanol and acetic acidunder gentle reflux, there is obtained methyl Nacety1-7-deoXy-7(S)-ethoxy-l-thio-u-lincosami nide (XI-II) as a syruphaving the molecular weight by mass spec. of 323 compared with thecalculated molecular weight of 323.41.

Part B-5: Methyl N-acetyl-2,3,4-tri-O-acetyl-7-deoxy-7(S)-ethoxy-a-thiolincosaminide (XIV) and methyl N- acetyl-7- deoxy-7(S) -ethoxy-2, 3 ,di-O-acetyl-u-thiolincosaminide (XV) On treating themethyl N-acety1-7-deoxy-7 (S)-eth0xy-athiolincosaminide (XIII) withacetic anhydride and pyridine by the process of Part A-3, there isobtained methyl N-acetyl-2,3,4-tri-O-acetyl-7-deoxy-7(S)-O-ethy1-mthiolincosaminide (XIV) together with a minor amount ofN-acetyl-2,3-di-O-acetyl-7 deoxy 7(S) ethoxy athiolincosaminide (XV).The products (isolated on a Craig in 500 transfers usingethanolzwaterzethyl acetatezcyclohexane (1:1:1:1) as the solvent system)are characterized as follows:

Mixture: M.P. 197-199" C. [d] +247 (c., 0.665, CHCl Analysis.-Calcd. forC H O NS (percent): C, 50.76; H, 6.95; N, 3.12; S, 7.13; OEt, 10.02.Found (percent): C, 50.42; H, 7.07; N, 3.18; S, 7.37; OEt, 11.85.

'Pure XIV (K=1.59): M.P. 254-255 C. [a] +199 (c., 0.8638, CHClAnalysis.Calcd. for C H O NS (percent): C, 50.76; H, 6.95; N, 3.12; S,7.13; OEt, 10.02. Found (percent): C, 50.75; H, 7.06; N, 3.37; S, 7.31;OEt, 10.25. Mol. wt. Calcd.: 449.52. Found (Mass spec.): 449.

Pure XV (K=0.87): M.P. 215.5-216.5 C. [a] +261 (c., 1.0448, CHClAnalysis.Calcd. for C H O NS (percent): C, 50.11; H, 7.17; N, 3.44; S,7.87. Found (percent): C, 50.17; H, 7.30; N, 3.50; S, 7.62. M01. wt.Calcd.: 407.48. Found (Mass spec.): 407.

'Part 06: Methyl 7-deoxy-7(S)-ethoxy-a-thiolincosaminide (XVI) NH2 HO \lj)/ SMa H XVI 0n subjecting the products of Part fB-5, that is, themixture, the pure XIV or the pure XV, to hydrazinolysis, there isobtained methyl 7-deoxy-7(S)-ethoxy-a-thiolincosaminide (XVI) having thefollowing characteristics:

M.P. 194-196 C. [a] I-252 (0., 0.7438, H O).

Analysis.Calcd. for C H O NS (percent): C, 46.95; H, 8.24; N, 4.98; S,11.40. Found (percent): C, 46.66; H, 8.09; N, 5.26; S, 11.33. Mol. wt.Calcd.: 281.37. Found (Mass spec.): 281.

Part D-S: 7-deoxy-7(S)-ethoxylincomyein hydrochloride (XVII) Me 1 1 on,

l-l -NH-- SMe 0H XVII On treating the methylN-acetyll6,7-aziridino-6-deamino-7-deoxy-a-thiolincosaminide (XII) withpropanol and acetic acid under gentle reflux, there is obtained methyl Nacetyl 7 deoxy 7 (S)-propoxy-u-thiolincosaminide (XVII) from which onacetylation with acetic anhydride in pyridine by the procedure of PartB-5, there is obtained methylN-acetyl-Z,3,4-tri-O-acetyl-7(S)-propoxy-7-deoxya-thiolincosaminide(XIX) containing a minor amount of methyl N acetyl 2,3 di-O-acetyl-7(S)-propoxy-7-deoxya-thiolincosaminide (XX) having the followingcharacteristics:

Mixture: M.P. 240-242 C. [a] +207 (c., 0.9054, CHCl Analysis.--Calcd.for C H 0 NS (percent): C, 51.81; H, 7.17; N, 3.03; S, 6.92. Found(percent): C, 51.41; H,

I SMe on XXI n hydrazinolysis of the above products (Part A-6) there isobtained methyl 7-deoxy-7(S)-propoxy-u-thiolincosaminide (XXI).

Part C-6: 7-deoxy-7(S)-propoxylincomycin hydrochloride (XXII) i N I CH3[6r x -OPr K .Hcl OH /L iii M.

H XXII On acylation withtrans-1-methyl-4-propyl-L-2-pyrroliline-carboxylic acid by the procedureof Part C-l, there is obtained 7-deoxy-7(S)-propoxylincomycinhydrochloride (XXII).

EXAMPLE 7 [Part A-7: Methyl N-acetyl-2,3,4-tri-O-acetyl-7-deoxy-7(S)-isopropoxy-u-thiolincosaminide (XXIII) O(iP r) AcNH- 0A0 XXIIIFollowing the procedure of Part B-2 but substituting the methanol byisopropyl alcohol, there is obtained methyl N acetyl2,3,4-tri-O-acetyl-7-deoxy-7(S)-isopropoXy-a-thiolincosarninide (XXIII)having the following characteristics: M.P. 253-254 C. [0:1 +192 (c.,0.5352, CHCl Analysis.Calcd. for C H O NS (percent): C, 51.81; H, 7.17;N, 3.03; S, 6.92. Found (percent): C, 51.96; H, 7.07; N, 3.19; S, 6.61.M01. Wt. Calcd.: 463.6. Found (Mass spec.): 463.

Part B-7: Methyl 7-deoxy-7(S)-isopropoxy-uthiolincosarninide (XXIV) OH lSMe OH XXIV On hydrazinolysis of compound XXIII (Part A-7), there isobtained methyl 7-deoXy-7(S)-isopropoxy-u-thiolinocosamiru'de having thefollowing characteristics: M.P. 2l2213 C. [a1 +225 (c., 0.376, H O).

Analysis.-Calcd. for C H O NS (percent): C, 48.79; H, 8.53; N, 4.74; S,10.86. Found (percent): C, 48.52; H, 8.55; N, 5.26; S, 10.84. Mol. Wt.Calcd.: 295.40. Found (Mass spec.): 295.

Part C-7: 7-deoxy-7 (S)-isopropoxylincomycin hydrochloride (XXV) -HClFollowing the procedure of Part C-l, compound XXV (Part C-7) isconverted to 7-deoxy-7(S)-isopropoxylincomycin hydrochloride having thefollowing characteristics: M.P. amorphous. [1x1 (c., 0.898, H O).

Analysis.-Calcd. for C -H O N S-HC1 (percent): C, 51.99; H, 8.52; N,5.78; S, 6.61; Cl, 7.31. Found (Corrected for 4.36% H O) (percent): C,51.72; H, 8.33; N, 5.59; S, 6.35; Cl, 7.29. M01. Wt. Calcd. (free base):448.62. Found: 448.

Activity: about the same as lincomycin.

EXAMPLE 8 Part A-S: Methyl N-acetyl-2,3,4-tri-O-acetyl-7-deoxy-(S)-cyclohexyloXy-a-thiolincosaminide (XXVI) O Cyclohexyl AcNH XXVI

Following the procedure of Part B-2 but substituting the methanol bycyclohexanol, there is obtained methyl N acetyl 2,3,4 tri O acetyl 7deoxy 7(S)- cyclohexyloxy a thiolincosaminide (XXVI) having thefollowing characteristics: M.P. 266268 C. [0:1 +163 (c., 1.0548, CHCIAnalysis.Calcd. for C H 0 NS (percent): C, 54.85; H, 7.41; N, 2.78; S,6.37. Found (percent): C, 54.93; H, 7.53; N, 2.87; S, 6.65. M01. Wt.Calcd.: 503.61. Found (Mass spec.): 503

Part B-8: Methyl 7(S)-cyclohexyloxy-7-deoxy-a thionlincosaminide (XXVII)-O Cyelohexyl \l l SMe 0H XXVII On hydrazinolysis of compound XXVI (PartA-8), there is obtained methyl 7 (S) cyclohexyloxy 7deoxya-thiolincosaminide (XXVII) 13 Part C-8:7(S)-cyclohexy1-7-deoxylincomycin hydrochloride (XXVIII) Following theprocedure of Part C-l, methyl 7 (S)- cyclohexyloxy 7 deoxy athiolincosaminide (XXVII) is converted to 7(S) cyclohexyloxy 7deoxylincomycin hydrochloride (XXVIII).

EXAMPLE 9 Part A-9: Methyl N acetyl 7 deoxy 7(S) 2' hydroxyethoxy athiolincosaminide (XXIX) and methyl N acetyl 2,3,4 tri O acetyl 7(S) 2'acetoxyethoxy-7-deoxy-tat-thiolincosaminide (XXX) Following theprocedure of Part A-l but substituting the methanol by 2 hydroxyethanol,there is obtained methyl N acetyl 7 deoxy 7(S) 2 hydroxyethoxyathiolincosaminide (XXIX) which when acylated by the procedure of PartA-3 but with heating on a steam bath to produce the fully acylatedproduct gives methyl N acetyl 2,3,4 tri O acetyl 7(S) 2 acetoxycthoxy 7deoxy a thiolincosaminide having the following characteristics: M.P.223225 C. [a] +172 (c. 1.0098, CHCl Analysis.Calcd. for C H O NS(percent): C, 49.69; H, 6.55; N, 2.76; S, 6.32. Found (percent): C,49.56; H, 6.63; N, 2.90; S, 6.63. M01. Wt. Calcd.: 507.55. Found: 507.

Part B-9: Methyl 7-deoxy-7(S)-2'-hydroxyethoxy-uthiolincosaminide (XXXI)CH2CH20H HO O K... M i...

XXXI

14 Part C-9: 7-Deoxy-7 (S)-2'-hydroxyethoxylincomycin hyrochloride Me cm6, --o0H.cH.oH

l I NH H01 6 no K... 1

l Me

OH XXXII Following the procedure of Part C-l, methyl 7-deoxy- 7 (S) 2'hydroxyethoxy a thiolincosaminide (XXXI) is converted to 7 deoxy 7(S) 2'hydroxyethoxylincomycin hydrochloride having the followingcharacteristics: M.P. amorphous 21 107 (c. 1.102, H O).

Analysis.Calcd. for C20H38O7N2S'HC]. (percent): C, 49.32; H, 8.07; N,5.75; S, 6.58; Cl, 7.28. Found (Corrected for 2.11% H O) (percent): C,49.61; H, 7.85; N, 5.54; S, 6.46; Cl, 7.76. M01. Wt. Calcd. (free base):450.59. Found (Mass spec.): 450. Activity: about /3 lincomycin.

EXAMPLE 10 Part A-lO: Methyl N acetyl 7 deoxy 7(S) 2'- methoxyethoxy athiolincosaminide (XXXIII) and N acetyl 2,3,4 tri O acetyl 7 deoxy 7(8)-2' methoxyethoxy-tat-thiolincosaminide (XXXIV) on, on,

-0CH CH OMe -OCHaCHzOMe AcNH- AeNH- HO A00 K0 H and 0 Ac l l SMe SMe 0H0A0 xxxrrr xxxrv Following the procedure of Part A-l but substitutingthe methanol by 2-methoxyethanol, there is obtained methylN-acetyl-7-deoxy-7(S)-2'-methoxyethoxy-a-thiolincosaminide (XXXII) whichon acetylation by the procedure of Part A-3 but with heating on a steambath to produce the fully acetylated product yields methylN-acetyl-2,3,4-tris O-acety1-7-deoxy 7(S)-2'-methoxyethoxy-ot-thiolincosaminide (XXXIV) which is characterizedas follows M.P. 222223 C. [(Z]D+177 (c. 1.0788, CHCl I Analysis.Calcd.for C H O NS (percent): C, 50.09; H, 6.94; N, 2.92; S, 6.69; OMe, 6.47.Found (percent): C, 50.13; H, 7.00; N, 2.77; S, 6.33; OMe, 7.28. M01.Wt. Calcd.: 497.54. Found (Mass spec): 479.

Part B-10: Methyl 7-deoxy-7(S)-2-methoxyethyltit-thiolincosaminide(XXXV) XXV 15 46.57; H, 8.32; N, 5.01; S, 10.70. Mol. Wt. Ca1cd.:311.40. Found (Mass spec.): 311.

Part C-lO: 7-deoxy-7 (S)-2'-methoxyethoxylincomycin hydrochloride Me lOH: -CHzCHz0Me i I HCl on I I/ SMe on XXXVI EXAMPLE 11 Methyl7-deoxy-7(S) hydroxy or thiolincosaminide (XXXVII) (methyl6-amino-6,8-dideoxy-L-threo-a-D- galacto-octopyranoside) on xxxvn PartA-ll: Methyl N-acetyl-7-deoxy-7(S)-hydroxy-athiolincosaminide (XXXVIII)(methyl 6-acetamido-6, 8-dideoxy-L-threo-a-D galacto-octopyranoside) ISMo H 'xxxvn'r To a solution of 2.35 gms. of methyl 6,7-aziridino-6-deamino-7-deoxy-a-thiolincosaminide (V) in 25 cos. of water was added2.04 grns. of acetic anhydride and the solution left at room temperatureovernight. The solution was then taken to dryness on a rotary evaporatorat 40 C./7 mm. to give a colorless syrup which was chromatographed on750 gms. of silica gel in a 4.8 x 98 cm. column using 1 MeOI-I27 CHCl asthe solvent system. After a forerun of 550 rnL, 50 ml. fractions werecollected. Fractions 90 through 160 were pooled and taken to dryness togive 2.3 gms. of methyl N-acetyl7-deoxy-7(S)-hydroxy-a-thiolincosaminide as a colorless solid whichcrystallized from methanol as colorless rods having the followingcharacteristics: M.P. 218-219 C. [d]n+260 (c., 1.0296, H O).

Analysis.-Calcd. for C H O NS (percent): C, 44.73; H, 7.17; N, 4.74; S,10.86. Found (percent): C, 44.89; H, 7.02; N, 5.16; S, 10.64. Mol. Wt.Calcd.: 295.36. Found (Mass spec.): 295.

Part B-ll: Deacetylation The crystallized material from Part A-ll wascombined with the mother liquors and taken to dryness on a rotaryevaporator at 40 C./ 7 mm. to give 201 gms. solid which was heatedovernight under gentle reflux with 40 cos. of hydrazine hydrate withstirring. The solvent was removed from the colorless solution on arotary evaporator at 7 mm. pressure in an oil bath at C. The resultingcolorless crystalline residue on recrystallization from methanol gavemethyl 7-deoxy-7(S)-hydroXy-a tl1iolincosaminide (XXXVII) as colorlessplatelets having the following characteristics: M.P. 211212 C. [Ot] +28O(c., 0.7728, H O).

Analysis.Calcd. for C H O NS (percent): C, 42.67; H, 7.56; N, 5.53; S,12.66. Found (percent): C, 42.81; H, 7.69; N, 5.85; S, 12.73. Mol. Wt.Calcd.: 253.32. Found (Mass spec.): 253.

EXAMPLE 12.

Methyl N acetyl-2,3,4-tri-O-acetyl-7"(S)-ethoxy-7-deoxyu-thiolincosaminide (XIV) and methyl N-ac etyl-2,3,4-

XIV

Following the procedure of Part B-.-2 but substituting the methanol byethanol there is obtained methyl N-acetyl-2,3,4-tri-O-acetyl-7(S)-ethoxy 7 deoxy-a-thiolincosaminide (XIV)identical with the product of Part B-5 and a minor amount of N-acetyl2,3,4-tri-O-acetyl7(S)- acetoxy 7 deoxy-a-thiolincosaminide- (XXXIX)which can be separated by Craig countercurrent distribution using 1EtOH:1 H O:1 EtOAc:1.5 cyclohexane as the solvent system in 500"transfers. The minor component '(XXXIX) was obtained from tubes numbers-200 (K=0.52), and the major component (XIV), fromttubes numbers 260-330(K'=1.43). The minor component (XXXIX) crystallized from ethylacetate ascolorless needles having the following characteristics: M.P. 312.- 313C. [a] +182'(c., 0.51898,'CHCl Analysis. Calcd. for C H O 'NS.(percent): C, 49.22; H, 6.31; N, 3.O2; S, 6.92, Found (percent): C,49.17; H, 6.51; N, 3.08; S, 6.81. M01. Wt. Calcd: 463.50. FoundKMassspec.): 463. g

On subjecting the minor component to hydrazinolysis there is obtainedmethyl 7 deoxy-'7'(S)-hydroxy-u-thiolincosaminide (XXXVIII) identicalwith the product of Part B-11.

I claim:

1. A process for making compounds of the formula AcNH- r Ae1T( 0101 i i1 l/ yl I OAcl j" -'1I r where Alkyl is alkyl of not more than 4c arbonatoms, .R is hydrogen or a hydrocarbon radical of not more, than 12carbon atoms or a hydroxy-substituted, alkoxy-substituted,orhalo-substituted hydrocarbon radical of not more than 12 carbon atoms,AC and AC are hydrogen or 17 acetyl which comprises opening theaziridino ring of a compound of the formula w-here Ac is acetyl andAlkyl and Ac are as given above by solvolysis with a compound of theFormula ROH wherein R is as given above and subjecting the product tohydrazinolysis when it is desired that Ac and Ac be hydrogen.

2. The process of claim 1 in which the solvolysis is effected in thepresence of acetic acid.

3. The process of claim 2 in which Ac is hydrogen.

4. The process of claim 2 in which Ac is acetyl.

5. The process of claim 3 in which ROH is lower alkanol.

6. The process of claim 4 in which ROH is lower alkanol.

7. The process of claim 2 in which a starting compound of the formula 7is N-acylated with acetic acid anhydride whereby acetic acid is freed tocatalyze the solvolysis to form a compound of the formula 10. Theprocess of claim 2 in which the starting compound has the formula HO O IS-Alkyl whereby the product has the formula AcN H- S-Alkyl wherein Ac isacetyl and R is lower alkyl and Alkyl is as given in claim 1.

18 11. The process of claim 2 wherein the starting compound has theformula whereby the product has the formula CHI . OAe

I l S-Alkyl 0A0 5 wherein Ac is acetyl, and R is lower alkyl and Alkylis as given in claim 1.

12. The process according to claim 2 in which a starting compound whereAc and Ac are hydrogen is peracetylated prior to the opening of theaziridino ring whereby a peracetylated product essentially free ofpartially acetylated product is obtained.

13. The process according to claim 2 in which the product isperacetylated after the aziridine ring is opened whereby a peracetylatedproduct is obtained concomitantly with a minor amount of a compound ofthe formula AcNH- OAc1 )L -Alkyl OAC1 wherein Ac and Ac are acetyl and-R and Alkyl are as given in claim 1.

14. The process according to claim 2 wherein the start- 5 ing compoundhas the formula CH; AcN

Ac O

OAGI i S-Alkyl A01 and the solvolysis is effected with ethanol andhigher alcohols whereby the product is obtained concomitantly with aminor amount of a compound of the formula wherein Ac, A0 and Ac areacetyl.

19 15. A compound of the formula I S-Alkyl wherein Ac is hydrogen oracetyl and Ac is hydrogen or acetyl, R is hydrocarbon containing notmore than 12 carbon atoms or hydroxy-substituted, alkoxy-substituted, orhalo-substituted hydrocarbon containing not more than 12 carbon atoms,and Alkyl is alkyl of not more than 4 carbon atoms.

19. The compound of claim 25 in which Ac is acetyl and Ac is hydrogen.

20. The compound of claim 18 in which Ac and A0 are acetyl.

21. The compound of claim 18 in which Ac and Ac are hydrogen.

22. The compound of claim 19 in which R is methyl, or acetoxyethyl.

23. The compound of claim 19 in which R is lower alkyl.

24. The compound of claim 21 in which R is methyl.

20 25. A compound of the formula AcNH- 0A0; wherein Ac and Ac are acetyland R and Alkyl is alkyl of not more than 4 carbon atoms.

26. A compound of the formula in which Ac, Ac and A0 are acetyl andAlkyl is alkyl of not more than 4 carbon atoms.

27. Methyl 7-O-methyl-6,8-dideoxy-6-(trans-l-methyl-4-propyl-L-2-pyrrolindinecarboxamido-l-thio-L-threo-a-D--galacto-octopyranoside) and the pharmacologically acid addition saltsthereof.

28. The hydrochloride salt of the compound of claim 27.

References Cited UNITED STATES PATENTS 3,300,475 1/1967 Bannister 260210R 3,326,891 6/1967 Hoeksema et al. 260-210 R 3,366,624 l/1968 Argoudeliset al. 260-210 R 3,544,551 12/1970 Kagan et al. 260-210 R LEWIS GOTTS,Primary Examiner J. R. BROWN, Assistant Examiner US. Cl. X.R. 260-25;999

